Use of Phospholipase A 2 for Antigen Retrieval in Zebrafish Whole-Mount Immunohistochemistry

Abstract

An advantage in zebrafish is that we can identify spatial and temporal patterns of protein expression using whole-mount immunohistochemistry. To allow primary antibodies to interact with their targets, most tissues must undergo some type of antigen retrieval. Many retrieval techniques have utilized protein-digesting enzymes to access antigens. Here we investigate the use of phospholipase A₂ (PLA₂) as the sole enzyme for antigen retrieval as well as in combination with low concentrations of proteinase K. Concentrations of proteinase K used with PLA₂ are unable to expose the antigen when used as the sole enzyme. We demonstrate that PLA₂ is useful for both nuclear and cytoplasmic antigens but not for extracellular matrix components.